NTA Chips for SPR: Difference between revisions
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Several parameters affecting level of ligand immobilization were studied: | Several parameters affecting level of ligand immobilization were studied: | ||
*conditions for NTA surface activation with Ni<sup>2+</sup>: [NiCl<sub>2</sub>], [NaCl] | *conditions for NTA surface activation with Ni<sup>2+</sup>: [NiCl<sub>2</sub>], [NaCl] and pH of activation buffer | ||
*effects of pH, NaCl and detergent (tween) | *effects of pH, [NaCl] and detergent (tween) concentration in the ligand immobilization buffer | ||
*effect of EDTA (0-300 mM, Na<sup>+</sup> salt) in the running and ligand buffers | *effect of EDTA (0-300 mM, Na<sup>+</sup> salt) in the running and ligand buffers | ||
*concentration of his-tagged ligand and number of his-tags per ligand molecule | |||
Also surface regeneration conditions were optimized. | |||
<biblio> | <biblio> | ||
#his1 pmid=9344407 | #his1 pmid=9344407 | ||
</biblio> | </biblio> |
Revision as of 19:26, 21 March 2008
Nieba et al. [1] investigated use of Ni2+-NTA chip surface for immobilization of his-tagged proteins.
Several parameters affecting level of ligand immobilization were studied:
- conditions for NTA surface activation with Ni2+: [NiCl2], [NaCl] and pH of activation buffer
- effects of pH, [NaCl] and detergent (tween) concentration in the ligand immobilization buffer
- effect of EDTA (0-300 mM, Na+ salt) in the running and ligand buffers
- concentration of his-tagged ligand and number of his-tags per ligand molecule
Also surface regeneration conditions were optimized.